LOCUS       sequence-131214-        8555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Sema4A protein, 
            C-terminally fused to alkaline phosphatase + 6X histidine tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8555)
  AUTHORS   Visser JJ, Cheng Y, Perry SC, Chastain AB, Parsa B, Masri SS, Ray 
            TA, Kay JN, Wojtowicz WM
  TITLE     An extracellular biochemical screen reveals that FLRTs and Unc5s 
            mediate neuronal subtype recognition in the retina.
  JOURNAL   Elife. 2015 Dec 3;4. pii: e08149. doi: 10.7554/eLife.08149.
  PUBMED    26633812
REFERENCE   2  (bases 1 to 8555)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2015-12-3"; volume: "4. pii: e08149. doi"; pages: " 
            10.7554/eLife.08149"
COMMENT     Sequence Label: Sema4a-AP-His
FEATURES             Location/Qualifiers
     source          1..8555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        25..408
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        409..612
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     563..583
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     609..633
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          748..1573
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
     regulatory      1604..1613
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5135..5152
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5178..5299
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5215..5234)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5269..5288
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5337..5354)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5508..6096)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5588..5607)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6267..7058,7059..7127))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6890..6909
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7128..7232)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7300..7318
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7356..7378)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7380..7400)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7385..7715
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7566..7701
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7628..7647
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7852..7871
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7938..8393)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8075..8096)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8287..8306
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8519..8541
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8533..8550
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8534..8550
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gtcgaggtct cgacggtatc gattgacatt gattattgac tagctagtta ttaatagtaa
       61 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      121 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      181 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      241 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt
      301 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac
      361 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      421 tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      481 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      541 cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      601 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
      661 gacctccata gaagacaccg ggaccgatcc agcctccgcg gccgggaacg gtgcattgga
      721 acgcggattc cccgtgccaa gagtgacgta agtaccgcct atagactcta taggcacacc
      781 cctttggctc ttatgcatgc tatactgttt ttggcttggg gcctatacac ccccgcttcc
      841 atatgctata ggtgatggta tagcttagcc tataggtgtg ggttattgac cattattgac
      901 cactccccta ttggtgacga tactttccat tactaatcca taacatggct ctttgccaca
      961 actatctcta ttggctatat gccaatactc tgtccttcag agactgacac ggactctgta
     1021 tttttacagg atggggtccc atttattatt tacaaattca catatacaac aacgccgtcc
     1081 cccgtgcccg cagtttttat taaacatagc gtgggatctc cacgcgaatc tcgggtacgt
     1141 gttccggaca tgggctcttc tccggtagcg gcggagcttc cacatccgag ccctggtccc
     1201 atgcctccag cggctcatgg tcgctcggca gctccttgct cctaacagtg gaggccagac
     1261 ttaggcacag cacaatgccc accaccacca gtgtgccgca caaggccgtg gcggtagggt
     1321 atgtgtctga aaatgagctc ggagattggg ctcgcaccgc tgacgcagat ggaagactta
     1381 aggcagcggc agaagaagat gcaggcagct gagttgttgt attctgataa gagtcagagg
     1441 taactcccgt tgcggtgctg ttaacggtgg agggcagtgt agtctgagca gtactcgttg
     1501 ctgccgcgcg cgccaccaga cataatagct gacagactaa cagactgttc ctttccatgg
     1561 gtcttttctg cagaagcttg aattctcgag gatccgcggc cgcgccacca tggccctacc
     1621 atccctgggc caggactcat ggagtctcct gcgtgttttt ttcttccaac tcttcctgct
     1681 gccatcactg ccacctgctt ctgggactgg tggtcagggg cccatgccca gagtcaaata
     1741 ccatgctgga gacgggcaca gggccctcag cttcttccaa caaaaaggcc tccgagactt
     1801 tgacacgctg ctcctgagtg acgatggcaa cactctctat gtgggggctc gagaggccgt
     1861 cctggccttg aatatccaga acccaggaat cccaaggcta aagaacatga taccctggcc
     1921 agccagtgag agaaaaaaga ccgaatgtgc ctttaagaag aagagcaatg agacacagtg
     1981 tttcaacttc attcgagtcc tggtctctta caatgctact cacctctatg cctgtgggac
     2041 ctttgccttc agccctgcct gtaccttcat tgaactccaa gattccctcc tgttgcccat
     2101 cttgatagac aaggtcatgg acgggaaggg ccaaagcccc tttgaccctg ttcacaagca
     2161 cacagctgtc ttggtcgatg ggatgcttta ttccggcacc atgaacaact tcctgggcag
     2221 cgagcccatc ctgatgcgga cactgggatc ccagcctgtt ctcaagactg acatcttctt
     2281 acgctggctg cacgcggatg cctccttcgt ggcagccatt ccatccaccc aggtcgtcta
     2341 tttcttcttt gaggagacag ccagcgagtt tgacttcttt gaagagctgt atatatccag
     2401 ggtggctcaa gtctgcaaga acgacgtggg cggtgaaaag ctgctgcaga agaagtggac
     2461 caccttcctc aaagcccagt tgctctgcgc tcagccaggg cagctgccat tcaacatcat
     2521 ccgccacgcg gtcctgctgc ccgccgattc tccctctgtt tcccgcatct acgcagtctt
     2581 tacctcccag tggcaggttg gcgggaccag gagctcagca gtctgtgcct tctctctcac
     2641 ggacattgag cgagtcttta aagggaagta caaggagctg aacaaggaga cctcccgctg
     2701 gaccacttac cggggctcag aggtcagccc gaggccaggc agttgctcca tgggcccctc
     2761 ctctgacaaa gccttgacct tcatgaagga ccattttctg atggatgagc acgtggtagg
     2821 aacacccctg ctggtgaagt ctggtgtgga gtacacacgg cttgctgtgg agtcagctcg
     2881 gggccttgat gggagcagcc atgtggtcat gtatctgggt acctccacgg ggtccctgca
     2941 caaggctgtg gtgcctcagg acagcagtgc ttatctcgtg gaggagattc agctgagccc
     3001 tgactctgag cctgttcgaa acctgcagct ggcccccgcc cagggtgcag tgtttgcagg
     3061 cttctctgga ggcatctgga gagttcccag ggccaattgc agtgtctacg agagctgtgt
     3121 ggactgtgtg cttgccaggg accctcactg tgcctgggac cctgaatcaa gactctgcag
     3181 ccttctgtct ggctctacca agccttggaa gcaggacatg gaacgcggca acccggagtg
     3241 ggtatgcacc cgtggcccca tggccaggag cccccggcgt cagagccccc ctcaactaat
     3301 taaagaagtc ctgacagtcc ccaactccat cctggagctg ccctgccccc acctgtcagc
     3361 actggcctct taccactgga gtcatggccg agccaaaatc tcagaagcct ctgctaccgt
     3421 ctacaatggc tccctcttgc tgctgccgca ggatggtgtc gggggcctct accagtgtgt
     3481 ggcgactgag aacggctact cataccctgt ggtctcctat tgggtagaca gccaggacca
     3541 gcccctggcg ctggaccctg agctggcggg cgttccccgt gagcgtgtgc aggtcccgct
     3601 gaccagggtc ggaggcggag cttccatggc tgcccagcgg tcctacggcg cgcctggatc
     3661 ttccggaatc atcccagttg aggaggagaa cccggacttc tggaaccgcg aggcagccga
     3721 ggccctgggt gccgccaaga agctgcagcc tgcacagaca gccgccaaga acctcatcat
     3781 cttcctgggc gatgggatgg gggtgtctac ggtgacagct gccaggatcc taaaagggca
     3841 gaagaaggac aaactggggc ctgagatacc cctggccatg gaccgcttcc catatgtggc
     3901 tctgtccaag acatacaatg tagacaaaca tgtgccagac agtggagcca cagccacggc
     3961 ctacctgtgc ggggtcaagg gcaacttcca gaccattggc ttgagtgcag ccgcccgctt
     4021 taaccagtgc aacacgacac gcggcaacga ggtcatctcc gtgatgaatc gggccaagaa
     4081 agcagggaag tcagtgggag tggtaaccac cacacgagtg cagcacgcct cgccagccgg
     4141 cacctacgcc cacacggtga accgcaactg gtactcggac gccgacgtgc ctgcctcggc
     4201 ccgccaggag gggtgccagg acatcgctac gcagctcatc tccaacatgg acattgacgt
     4261 gatcctaggt gggggccgaa agtacatgtt tcgcatggga accccagacc ctgagtaccc
     4321 agatgactac agccaaggtg ggaccaggct ggacgggaag aatctggtgc aggaatggct
     4381 ggcgaagcgc cagggtgccc ggtacgtgtg gaaccgcact gagctcatgc aggcttccct
     4441 ggacccgtct gtggcccatc tcatgggtct ctttgagcct ggagacatga aatacgagat
     4501 ccaccgagac tccacactgg acccctccct gatggagatg acagaggctg ccctgcgcct
     4561 gctgagcagg aacccccgcg gcttcttcct cttcgtggag ggtggtcgca tcgaccatgg
     4621 tcatcatgaa agcagggctt accgggcact gactgagacg atcatgttcg acgacgccat
     4681 tgagagggcg ggccagctca ccagcgagga ggacacgctg agcctcgtca ctgccgacca
     4741 ctcccacgtc ttctccttcg gaggctaccc cctgcgaggg agctccatct tcgggctggc
     4801 ccctggcaag gcccgggaca ggaaggccta cacggtcctc ctatacggaa acggtccagg
     4861 ctatgtgctc aaggacggcg cccggccgga tgttaccgag agcgagagcg ggagccccga
     4921 gtatcggcag cagtcagcag tgcccctgga cgaagagacc cacgcaggcg aggacgtggc
     4981 ggtgttcgcg cgcggcccgc aggcgcacct ggttcacggc gtgcaggagc agaccttcat
     5041 agcgcacgtc atggccttcg ccgcctgcct ggagccctac accgcctgcg acctggcgcc
     5101 ccccgccggc accaccgacg ccgcgcaccc gggtcatcat caccatcacc attaatctag
     5161 agatcttatt aaagcagaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5221 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5281 aactcatcaa tgtatcttat catgtctggt cgactctagc tagactcttc cgcttcctcg
     5341 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5401 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5461 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5521 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5581 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5641 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5701 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5761 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5821 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5881 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5941 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6001 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6061 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6121 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6181 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6241 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6301 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6361 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6421 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6481 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6541 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6601 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6661 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6721 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6781 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6841 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6901 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6961 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7021 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7081 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7141 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7201 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7261 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     7321 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     7381 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc
     7441 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     7501 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     7561 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     7621 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     7681 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     7741 ttttcgtccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     7801 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     7861 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     7921 gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     7981 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     8041 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     8101 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     8161 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     8221 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     8281 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     8341 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg
     8401 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     8461 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     8521 cagtcacgac gttgtaaaac gacggccagt gaatt
//
